CN110772558A - Platycodon grandiflorum decoction extract and preparation method and application thereof - Google Patents

Abstract

The invention discloses an extract of Campanulaceae decoction and a preparation method and application thereof, belonging to the field of traditional Chinese medicine chemistry. The preparation method comprises: mixing Campanulaceae and licorice, crushing and soaking to obtain a mixed medicine; heating and refluxing an aqueous solution of the mixed medicine, Filter and collect the filtrate; after the filtrate is adsorbed by non-polar macroporous adsorption resin, the eluent is eluted to obtain an eluent; acetonitrile and water are used as mobile phases, and the concentrated eluent is analyzed in liquid chromatography. Gradient elution separation was performed, 30%-33% acetonitrile aqueous solution was collected, concentrated, and freeze-dried to obtain the Bellflower Decoction extract. The Bellflower Decoction extract includes licorice chalcone B, glycyrrhizin G2 and Ural licorice A; the Bellflower decoction extract has the effect of inhibiting the activity of Staphylococcus aureus. The invention adopts operations such as water extraction, non-polar macroporous resin purification, grading elution with different alcohol concentrations, etc., the operation method is simple, and the realization is strong, and the obtained Campanulaceae decoction extract can be used in anti-Staphylococcus aureus medicines.

Description

A kind of Campanulaceae decoction extract and its preparation method and application

technical field

The invention relates to the field of traditional Chinese medicine chemistry, in particular to an extract of Bellflower Decoction, a preparation method thereof, and an application in inhibiting the activity of Staphylococcus aureus.

Background technique

Acute lung injury (ALI) refers to the acute hypoxic respiratory insufficiency syndrome caused by infection or trauma to the body, resulting in the destruction of the pulmonary endothelial and epithelial cell barriers, resulting in diffuse interstitial diffusion and pulmonary edema.

Bacterial infection is a common cause of acute lung injury. There are two main forms of acute lung injury caused by bacterial infection: one is Gram-negative bacilli infection, such as Klebsiella pneumoniae, and LPS-induced ALI animal models are often used in experiments. Simulates a gram-negative bacillus infection; the other is a gram-positive coccus infection, such as Staphylococcus aureus, an important pathogen that causes infectious pneumonia; with the increasing resistance of Staphylococcus aureus, the The isolation rate of oxycillin Staphylococcus aureus (MRSA) has reached more than 60%, and the isolation rate of community-acquired MRSA (CA-MRSA) has also increased in recent years; because most MRSA strains are multi-drug resistant, they are not only resistant to almost all β-lactams. Antibiotic resistance, and high resistance to other antibacterial drugs such as fluoroquinolones, aminoglycosides, etc., has become one of the problems in clinical anti-infective treatment.

Infectious diseases belong to the category of warm and febrile diseases in the theory of traditional Chinese medicine. The treatment with traditional Chinese medicines of clearing heat and detoxification has a good effect. A large number of modern studies have shown that the chemical components of traditional Chinese medicines have good antibacterial activity. Combination of antibiotics can reverse the advantages of antibiotic resistance and low adverse reactions. Finding natural antibiotics has become a new way to solve bacterial resistance.

Campanulaceae Tang is composed of two traditional Chinese medicines, Campanulaceae and Licorice. It was originally from Zhang Zhongjing's "Treatise on Febrile Diseases: Shaoyin": "Shaoyin disease, two or three days, sore throat can be used with Gancao Tang; those who do not suffer from malnutrition, can be combined with Campanulaceae Tang." It is also recorded in "Golden Chamber Synopsis: Lung Fistula Lung Carbuncle Cough": "Coughing and chest fullness, vibrating cold pulse, dry throat but not thirsty, sometimes turbid saliva and fishy smell, long-term vomiting of pus like rice porridge, it is lung carbuncle, Campanulaceae decoction The main thing." Platycodon grandiflorum is hard and flat, can disperse the lungs and pharynx, expel phlegm and pus; licorice is sweet, can invigorate the spleen and replenish qi, clear heat and detoxify, expel phlegm and relieve cough, relieve acute pain and relieve pain. The combination of the two is used to treat sore throat and lung abscess cough.

Publication No. CN101396425A discloses a preparation method of a Bellflower Decoction preparation. In order to return to the essence of traditional medicine for treating diseases, it is proposed to extract the medicine with water according to the guiding ideology of "Zhou Gu", and combine modern concentration and preparation methods. It summarizes the preparation process parameters and applicable excipients that best match the Bellflower Decoction preparation; it describes the preparation method of the Bellflower Decoction, but does not involve the application of the Bellflower Decoction in inhibiting the activity of Staphylococcus aureus.

Publication No. 101396425A is a kind of traditional Chinese medicine for the treatment of chronic pharyngitis, and it is made into oral lozenges. Campanulaceae and licorice are Chinese medicinal materials of the same origin as medicine and food (traditional Chinese medicine that can be used for health care). Formed into buccal tablets, it has obvious effects of clearing away heat and detoxification, soothing throat and relieving pain, and has good preventive and therapeutic effects on various syndromes of Shaoyin disease and sore throat; it is mainly aimed at the treatment of pharyngitis with Bellflower Decoction, and does not involve LPS-induced acute Lung injury is protective.

SUMMARY OF THE INVENTION

The invention provides a preparation method of a Bellflower Decoction extract. In the preparation method, operations such as water extraction, non-polar macroporous resin purification, grading elution with different alcohol concentrations and the like are adopted to obtain the Bellflower Decoction with stronger activity of functional components. Extract.

A preparation method of bellflower soup extract, comprising:

(1) pulverizing, soaking after mixing Platycodon grandiflorum and licorice, obtain mixed medicine;

(2) the aqueous solution of the mixed medicine is heated to reflux, filtered, and the filtrate is collected;

(3) using non-polar macroporous adsorption resin to carry out adsorption to the filtrate of step (2) and eluting with eluent to obtain eluent;

(4) Using acetonitrile and water as mobile phases, the concentrated eluent is subjected to gradient elution separation in liquid chromatography, and the separated liquid is collected and concentrated to obtain an extract of Bellflower Decoction.

In step (1), the mass ratio of described Platycodon grandiflorum and licorice is 1:1-5.

In order to better dissolve the medicinal components in Platycodon grandiflorum and licorice in the solvent, the soaking solvent is water, the soaking time is 2-24h, and the soaking temperature is 25-50°C.

In step (2), the temperature of the heating and refluxing is 110-150° C., and the heating and refluxing time is 1-12 h.

In order to fully dissolve the medicinal components in Platycodon grandiflorum and licorice root in water, in step (2), after the mixed medicine is filtered, the medicinal residues are added to the water again, and the heating and refluxing times are 2-4 times.

In step (3), the non-polar macroporous adsorption resin is one of D101, XDA-5 or HP-10; because D101 macroporous resin has a considerable specific surface area and suitable pore size, The saponins have good selectivity, preferably, the macroporous adsorption resin is D101.

The adsorption time of the macroporous adsorption resin to the filtrate is 2-5h, and the medicinal components of Platycodon grandiflorum and licorice in the filtrate are absorbed more completely.

Due to the high polarity of the ethanol aqueous solution, the adsorbed drug can be completely eluted without damaging the macroporous resin. The eluent is 20%-40% ethanol aqueous solution, preferably 40%. % ethanol in water.

In step (4), the concentrated eluent is subjected to gradient elution separation in liquid chromatography, and the separation solution is collected, and the separation solution is a 30%-33% acetonitrile aqueous solution.

The present invention also provides a Bellflower Decoction extract prepared by the above preparation method. The Bellflower Decoction extract comprises Licorice Chalcone B, Glycyrrhizin G ₂ , and Ural Licorice A; B) As the main component of the Bellflower Decoction extract, the content is 50-60%.

The present invention also provides the application of the above-mentioned Bellflower Decoction extract in inhibiting the activity of Staphylococcus aureus.

The beneficial effects of the present invention are:

(1) operations such as water extraction, non-polar macroporous resin purification, and different alcohol concentration fractional elution are adopted in the preparation method of the Bellflower Decoction extract of the present invention, the operation is simple, the realization is strong, and the active ingredient activity Strong bellflower soup extract.

(2) The Bellflower Decoction extract of the present invention includes licorice chalcone B, glycyrrhizin G2, and Ural licorice A; wherein licorice chalcone B is the main component of the bellflower decoction extract, with a content of 50-60%.

(3) In the present invention, it is proved by the animal test research and the antibacterial activity experimental research of the Bellflower Decoction Extract that the Bellflower Decoction Extract has the effect of inhibiting the activity of Staphylococcus aureus.

Description of drawings

Fig. 1 is the chromatogram of HPLC analysis of the D1 extract of Bellflower Decoction in Example 1 of the present invention, wherein 1 is licorice chalcone B, 2 is glycyrrhizin G2, and 3 is ural licorice A.

Fig. 2 is the mass spectrum of licorice chalcone B in the extract D1 of Bellflower Decoction in Example 1 of the present invention.

Figure 3 is the mass spectrum of Glycyrrhizin G2 in the extract D1 of Bellflower Decoction in Example 1 of the present invention.

Fig. 4 is the mass spectrum of Ural licorice A in the extract D1 of Bellflower Decoction in Example 1 of the present invention.

Fig. 5 is a diagram showing the effect of the Kikyeng decoction on the pathological morphology of lung tissue in the present invention; wherein, A is the lung tissue section of the male rat in the blank control group; B is the lung tissue section of the male rat in the model group; C is the male rat in the non-infected group with Kikyeng Decoction Lung tissue slices, D is the lung tissue slices of male rats in the Kikyeng decoction infection group.

Figure 6 is a statistical diagram of bacterial load in the lungs of male mice in the present invention; Figure A is a statistical diagram of CFU results of male rat lung homogenate; Figure B is a statistical diagram of CFU results of bronchoalveolar lavage fluid.

Detailed ways

The present invention further describes the essential content of the present invention in detail with reference to the accompanying drawings and the following embodiments, which are only used to illustrate the present invention and not to limit the present invention.

Example 1

(1) 66.6g of Platycodon grandiflorum and 133.3g of licorice were pulverized and mixed, and 1.2L of water was added to soak for 12h to obtain a medicinal material mixed solution;

(2) Put the mixed solution of medicinal materials at 120°C for reflux extraction for 3 hours, and filter; add 0.8L of water to the filter residue and reflux at 120°C for 3 hours, combine the extracts, and concentrate under reduced pressure to obtain the total extract A1;

(3) Take 50mL of the total extract, put it on a packed D101 macroporous resin chromatographic column (4.6cm×30cm), and let it stand for adsorption for 3h; elute with 2.4L of 20% and 40% ethanol aqueous solution by volume respectively, Collect 40% ethanol-water eluted fractions;

(4) The 40% ethanol water eluate was concentrated to 30 mL under reduced pressure, 4 mL of the concentrated solution was taken and separated by liquid chromatography, 30%-33% acetonitrile water eluate was collected, concentrated, and freeze-dried to obtain 16 mg of Platycodon grandiflorum extract. object D1;

The conditions for liquid chromatography separation are: Agilent Zorbax SB-C18 column (21.2mm×250mm, 7μm); mobile phases are water (phase A) and acetonitrile (phase B); flow rate is 10 mL/min; gradient elution The program is 0min, 15%B; 40min, 30%B; 55min, 45%B; 60min, 80%B; 65min, 100%B; 70min, 100%B; UV detection wavelength 254nm.

Antibacterial activity evaluation of the extracts A1 and D1 of Bellflower Decoction

Concentrating and freeze-drying the extract in step (2) of Example 1 to obtain the total extract A1 of Campanulaceae decoction without purification and separation and the extract D1 of Campanula officinalis decoction obtained in step (4) as the experimental group, respectively weighing etc. Amounts of A1 and D1, dissolved in DMSO, were prepared as 100 mg/ml stock solutions, respectively. In bacterial incubation experiments, the stock solutions of A1 and D1 were diluted to 1 mg/ml with TSB medium, respectively.

Pick up Staphylococcus aureus monoclonal colonies from the blood plate and add them to a centrifuge tube containing 10ml of TSB medium, and in a constant temperature shaker at 37°C and 200rpm overnight for a total of 12h; take 100μl of bacterial solution the next day and add it to 10ml of fresh TSB medium 4 hours later, the bacteria were shaken at 2600rpm×10min for centrifugation, the supernatant was discarded, and phosphate buffered saline (PBS) was added to dilute to a concentration of 10 ⁸ CFU/ml; further dilute to 10 ⁴ CFU/ml with TSB medium as a backup bacterial liquid;

Take a 96-well plate, add 190 μl of the prepared A1 or D1 stock solution and 10 μl bacterial solution to each well, so that the bacterial solution in each well is 100 CFU, as the experimental group; take a 96-well plate, add 190 μl TSB to each well for culture Base and 10 μl of bacterial liquid were used as control group; then three groups of well plates were placed in a 37 °C bacterial incubator for 16 h.

The bacterial solution in the 96-well plate was pipetted evenly. After 10-fold dilution of the bacterial solution with PBS, 8 gradients were taken, and 5 μl of each was placed on the plate. After 24 hours of incubation in a bacterial incubator, CFU was counted, and the average value of three wells was taken. The experiment was repeated. Three times, the result is the average of three times;

Bacteriostatic rate = (control group CFU - experimental group CFU)/control group CFU × 100%

The results showed that the bacteriostatic rate of total extract A1 was (68±28)%, and the bacteriostatic rate of extract D1 was (100±0)%. Compound D1 showed strong antibacterial activity against Staphylococcus aureus.

HPLC-MS analysis of the extract D1 of Bellflower Decoction

Weigh the extract D1 of Bellflower Decoction, dissolve it in 50% methanol-water solution, and prepare a solution of D1 solution of Bellflower Decoction with a concentration of 1 mg/mL. After filtering, transfer it to a sample bottle for HPLC chromatographic analysis. The analysis results are shown in Figure 1. (1 is Glycyrrhizic Chalcone B, 2 is Glycyrrhizin G ₂ , and 3 is Ural Glycyrrhiza A), which shows that the main component in Campanulaceae extract D1 is Glycyrrhizic Chalcone B, and the content is 54%; Figure 2 is the mass spectrum of Glycyrrhiza chalcone B; the Campanulaceae decoction extract D1 also includes Glycyrrhizin G ₂ and Ural Glycyrrhiza A, wherein the mass spectrum of Glycyrrhizin G2 is shown in Figure 3; Mass spectrum of Ural licorice A.

HPLC chromatographic analysis conditions: Zorbax SB-C18 rapid analysis column (4.6mm×50mm, 1.8μm) from Agilent, USA; mobile phase: 0.05% formic acid water (phase A) and acetonitrile (phase B); flow rate: 0.6mL/min ; column temperature is 30°C; linear gradient: 0 min, 6% B; 35 min, 40% B; 60 min, 100% B; 70 min, 100% B; injection volume: 10 μL; detection wavelength: 254 nm.

Detection of minimum inhibitory concentration and minimum bactericidal concentration of licorice chalcone B

The minimum inhibitory concentration of licorice chalcone B was detected by the micro broth dilution method, and the minimum bactericidal concentration was detected:

(1) take by weighing licorice chalcone B and antibiotics (oxacillin sodium, vancomycin hydrochloride, chloramphenicol, streptomycin sulfate) respectively, prepare the drug stock solution of 12.8mg/ml with DMSO respectively;

(2) Add 100 μl of TSB medium to each well of the 96-well plate; add the sample to be tested (4 μl of drug stock solution + 96 μl of TSB medium) to the first column of wells, and add 100 μl from the first well to the second well, Dilute in half to the 11th column, and the 12th column is the control well;

(3) Add 100 μl (1×10 ⁵ CFU/ml) of the diluted standby bacterial solution to each well, at this time, the concentration of the drug stock solution from the first column to the eleventh column is 128 μg/ml, 64 μg/ml, 32 μg/ml, respectively. ml, 16μg/ml, 8μg/ml, 4μg/ml, 2μg/ml, 1μg/ml, 0.5μg/ml, 0.25μg/ml and 0.125μg/ml;

(4) Put the 96-well plate into the bacterial incubator and incubate for 24 hours, then take it out for observation; no bacterial growth can be seen with the naked eye, and the minimum drug concentration in the well-cleaned culture solution is the minimum inhibitory concentration (MIC) of the drug;

(5) Take 10 μl of liquid from all antibacterial wells and spread them on TSB agar plates in the original order, incubate at 37°C for 24 hours; the lowest drug concentration that does not show bacterial growth to the naked eye is the minimum bactericidal concentration (MBC).

Table 1 MIC and MBC results of licorice chalcone B and antibiotics

As shown in Table 1, the minimum inhibitory concentration and the minimum bactericidal concentration of licorice chalcone B and antibiotics were detected by the micro-broth dilution method. The minimum inhibitory concentration of licorice chalcone B was 4-8 μg/ ml; the minimum bactericidal concentration is 128 μg/ml.

Animal experiments on the treatment of Staphylococcus aureus pneumonia with the Simulated Bellflower Decoction Extract A1

In order to better simulate clinical Staphylococcus aureus pneumonia, the present invention uses airway instillation of Staphylococcus aureus to induce acute lung injury model in male rats for the first time to study the anti-S.

animal grouping

(1) C57BL/6 female mice aged 6-8 weeks, weighing 20g, were randomly divided into 4 groups, namely blank control group, model group, non-infected group with Bellflower Decoction, and infected group with Bellflower Decoction. They were adaptively reared in the laboratory before the experiment. One week, during which a normal diet was given, fasting for 12 hours before the experiment, and normal water supply;

(2) Blank control group: male rats were given 0.2 ml of distilled water; model group: 0.2 ml of distilled water was given to male rats, and 50 μl of Staphylococcus aureus solution (5×10 ⁷ CFU) was instilled into the airway after 2 hours; Platycodon grandiflorum Tang-uninfected group: male rats were given 0.2ml of Bellflower Decoction Extract A1 (2.7g/kg); Campanulaceae Decoction infected group: 0.2ml of Bellflower Decoction Extract A1 (2.7g/kg) was given to male rats by gavage, 2 hours later 50 μl of Staphylococcus aureus solution (5×10 ⁷ CFU) was instilled into the airway.

HE staining of histopathological sections of rat lungs

(1) Material sampling: The male mice were divided into 4 groups according to the above. After 24 hours of infection, they were killed by cervical dislocation, the thoracic cavity was opened, the lungs were taken and placed in 4% paraformaldehyde to fix for more than 24 hours, and the lung tissue was trimmed and flattened;

(2) Dehydration: dehydrate the lung tissue in sequence with 75% alcohol for 4 hours; 85% alcohol for 2 hours; 90% alcohol for 2 hours; 95% alcohol for 1 hour; anhydrous ethanol for 30 minutes, twice in a row; alcohol and benzene for 10 minutes; Xylene dehydration for 10min, twice in a row; wax soaking in 1h for three consecutive times;

(3) Embedding: the pulmonary tissue soaked in wax is embedded in an embedding machine, taken out and cooled in a -20° refrigerator to obtain a wax block for embedding the pulmonary tissue;

(4) Sectioning: place the wax block obtained in step (3) on a paraffin microtome to section (about 4 μm in thickness), and bake the slices in a 60° C. oven to obtain a paraffin section containing lung tissue;

(5) Dewaxing of paraffin sections: immerse the paraffin sections in xylene for 20 minutes, repeat twice; anhydrous ethanol for 10 minutes, repeat twice; 95% alcohol for 5 minutes; 90% alcohol for 5 minutes; 80% alcohol for 5 minutes; 70% alcohol for 5 minutes ;Final distilled water wash;

(6) Hematoxylin staining of nuclei: Harris hematoxylin staining for 5 min, rinsed with distilled water; differentiated with 1% hydrochloric acid alcohol for several seconds, rinsed with distilled water; 0.6% ammonia water returned to blue, rinsed with distilled water;

(7) Eosin-stained cytoplasm: eosin staining solution for 2 min;

(8) Dehydration and sealing: put the paraffin sections into 95% alcohol for 5 min twice, absolute ethanol for 5 min twice, xylene for 5 min twice, and finally seal the paraffin section containing lung tissue with neutral gum.

Statistics of bacterial load in the lungs of male mice

(1) Lung homogenization: The male mice were divided into 4 groups according to the above. After 24 hours of infection, they were sacrificed by cervical dislocation. The chest cavity was opened to take the lungs, which were placed in a QSP tube containing 1 ml of sterile PBS, and magnetic beads were placed in the homogenate. Grind in the machine at 65 Hz for 250 s until there are no large pieces of tissue; take 20 μl of tissue homogenate into a 96-well plate, add 180 μl of sterile PBS, dilute 10 times in 4 gradients, spot plate on TSB agar plate, and place in a bacterial incubator overnight. The number of colonies was counted the next day.

(2) Bronchoalveolar lavage fluid: The male mice were divided into 4 groups according to 2.2.2. After 24 hours of infection, they were anesthetized with 1.5% sodium pentobarbital intraperitoneally. Make an incision and insert a cannula; take 1ml of PBS back and forth for lavage twice, the recovery rate is over 80%, and collect it in a 2ml EP tube; take 20μl of bronchoalveolar lavage fluid into a 96-well plate, add 180μl of sterile PBS, and dilute 10 times 4 gradients were spotted on TSB agar plates, overnight in a bacterial incubator, and CFU was counted the next day.

Effects of Kikyo Decoction on Pathological Morphology of Lung Tissue

As shown in Figure 5, the effect of Kikyeng Decoction on the pathological morphology of lung tissue, A is the lung tissue section of the male rat in the blank control group, B is the lung tissue section of the male rat in the model group, and C is the lung tissue of the male rat in the non-infected group with Kikyeng Decoction Section picture, D is the section picture of the lung tissue of the male rat in the Kikyeng decoction infection group; the lung tissue of the male rat in the blank control group was normal, with abundant alveoli and intact bronchial structure (as shown in Figure 5A), acute lung injury induced by Staphylococcus aureus Pathological changes are often manifested as a large number of neutrophil infiltration.

In the experimental model group, more neutrophils were infiltrated in the blood vessels and outside the bronchi of the lung tissue of the male mice, as shown by arrow 1; the alveolar wall was thickened, and neutrophils were scattered in it, as shown by arrow 2; Red blood cells can be seen in the blood vessel, as shown by arrow 3; lymphocytes can be seen in the lumen of the blood vessel, as shown by arrow 4, these changes are highly similar to the pathological damage of ALI, which better simulates the clinical pathology of ALI caused by Staphylococcus aureus change (as shown in Figure 5B).

The lung tissue of male rats in the non-infected group with Kikyeng decoction was normal, with abundant alveoli and intact bronchial structure (as shown in Figure 5C), indicating that Kikyeng decoction would not affect the lung tissue of male rats.

Part of the blood vessels in the lung tissue of male rats in the Kikyeng decoction group were congested, as shown by the black arrows; there were no other obvious abnormalities in the tissue, the number of alveoli was abundant, and the bronchial structure was intact (as shown in Figure 5D). Histopathological morphology of rat lung tissue.

Effect of Kikyo Decoction on Lung Bacteria Loading

As shown in Figure 6, the amount of bacteria in the lungs is a direct indicator for testing the antibacterial effect of drugs. After 24 hours of infection, the lung tissue of the male mice in the model group and the Bellflower Decoction infection group was taken to make a homogenate, and a part was taken for dilution and spotting, as shown in the figure As shown in 6A, the CFU results of lung homogenate showed that the bacterial load in the lungs of the Bellflower Decoction infection group was significantly lower than that of the model group. One male rat died. As shown in Figure 6B, the CFU results of the bronchoalveolar lavage fluid showed that the bacterial load in the bronchoalveolar lavage fluid of the male rats in the Bellflower Decoction infection group was significantly lower than that in the model group; the above results show that the Bellflower Decoction can effectively reduce the The bacterial load in the lungs of male mice infected with bacteria has antibacterial effect in vivo.

The above results showed that Kikyo Decoction can effectively reduce the bacterial load in the lungs, improve the pathological morphology of lung tissue, and has a therapeutic effect on the lung injury caused by Staphylococcus aureus.

Claims (8)

1. a preparation method of Platycodon grandiflorum decoction extract, is characterized in that, comprises the following steps: (1) pulverizing, soaking after mixing Platycodon grandiflorum and licorice, obtain mixed medicine; (2) the aqueous solution of the mixed medicine is heated to reflux, filtered, and the filtrate is collected; (3) after the filtrate of step (2) is adsorbed by the non-polar macroporous adsorption resin, the eluent is eluted to obtain an eluent; (4) Using acetonitrile and water as mobile phases, the concentrated eluent is subjected to gradient elution separation in liquid chromatography, 30%-33% acetonitrile aqueous solution is collected, concentrated, and freeze-dried to obtain an extract of Bellflower Decoction. 2. the preparation method of bellflower soup extract according to claim 1, is characterized in that, in step (1), the mass ratio of described bellflower root and licorice is 1:1-5. 3 . The preparation method of Platycodon grandiflorum soup extract according to claim 1 , wherein, in step (2), the temperature of the heating and refluxing is 110-150° C., and the time of heating and refluxing is 1-12 h. 4 . 4. The preparation method of Platycodon grandiflorum extract according to claim 1, is characterized in that, in step (3), described non-polar macroporous adsorption resin is D101, XDA-5 or HP-10. 5. The preparation method of Platycodon grandiflorum soup extract according to claim 1, is characterized in that, in step (3), described adsorption time is 2-5h. 6. The preparation method of Platycodon grandiflorum soup extract according to claim 1, is characterized in that, in step (3), described eluent is the ethanol aqueous solution of 20%-40%. 7. A Bellflower Decoction extract prepared by the preparation method according to any one of claims 1-6. 8. a kind of application of bellflower soup extract according to claim 7 in inhibiting the activity of Staphylococcus aureus.

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